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Impact of miR-7 over-expression on the proteome of Chinese hamster ovary cells

Paper ID Volume ID Publish Year Pages File Format Full-Text
23868 43478 2012 12 PDF Available
Title
Impact of miR-7 over-expression on the proteome of Chinese hamster ovary cells
Abstract

MicroRNAs play critical roles in the regulation of biological processes such as growth, apoptosis, productivity and secretion thus representing a potential route toward enhancing desirable characteristics of mammalian cells for biopharmaceutical production. We have previously found that miR-7 over-expression significantly inhibits the growth of CHO-SEAP cells without impacting cellular viability, with an associated increase in normalised productivity. Understanding the biological basis of this effect might open the way to new strategies for bioprocess-relevant growth regulation. In this study we have carried out a quantitative label-free LC–MS profiling study of proteins exhibiting altered levels following over-expression of miR-7 to gain insights into potential mechanisms involved in the observed phenotype.From the analysis we found 93 proteins showing decreased levels and 74 proteins with increased levels following over-expression of miR-7. Pathway analysis suggests that proteins involved in protein translation (e.g. ribosomal proteins), RNA and DNA processing (including histones) are enriched in the list of proteins showing decreased expression. Proteins involved in protein folding and secretion were found to be up-regulated following miR-7 over-expression. In silico bioinformatic analysis using miRWalk, which combined the output from 6 selected miRNA target prediction algorithms, was used to evaluate if any of the down-regulated proteins were potential direct targets of miR-7. Two genes, stathmin and catalase, which both have known roles in the regulation of cellular growth, were found to overlap a number of the predictive target database searches in both mouse and rat, and are likely to be possible direct targets of miR-7 in CHO cells. This is the first report investigating the impact of a miRNA on the proteome of CHO cells.

► MiR-7 alters the growth of recombinant Chinese hamster ovary cells. ► First study on effects of dysregulating miRNA expression on the proteome of CHO cells. ► Label-free LC–MS identifies 168 altered proteins following miR-7 over-expression. ► Stathmin and catalase identified as possible direct targets of miR-7 in CHO cells.

Keywords
CHO, Chinese hamster ovary; SEAP, secreted alkaline phosphatase; LC–MS, liquid chromatography–mass spectrometry; RT, retention time; mgf, MASCOT generic filemicroRNA; miR-7; Label-free LC–MS proteomics; Chinese hamster ovary; Cell growth
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Impact of miR-7 over-expression on the proteome of Chinese hamster ovary cells
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 160, Issues 3–4, 31 August 2012, Pages 251–262
Authors
, , , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us